Tags

Type your tag names separated by a space and hit enter

DNA elements and protein factors involved in the transcription of the beta 2-adrenergic receptor gene in rat liver. The negative regulatory role of C/EBP alpha.
Biochemistry. 1996 Oct 08; 35(40):13136-46.B

Abstract

Primer extension and RNase protection analyses of the rat beta 2-adrenergic receptor (beta 2AR) gene identify two transcription start points at -64 and -220 nt, respectively. Transient transfections of putative promoter/pCAT constructs into DDT1 MF-2 cells indicate that fragments -36 to -100 (PI) and -186 to -312 (P2) are sufficient to promote transcription, whereas -911 to -1122 contains a negative regulatory element(s). RNase protection analysis of the 3' untranslated region (3'-UTR) indicates the presence of two transcripts with 3'-UTR of 111 and 604 nt exclusive of the poly(A+) tails. Northern blots of beta 2AR mRNA using full-length and partial cDNA probes indicate that a major 2.2 kb and a minor 1.6 kb species arise from the use of alternative promoters as well as different polyadenylation signals. DNase I footprinting and DNA mobility shift assays (DMSA) using rat liver nuclear extracts identify a number of transcription factors binding to sequence elements within or upstream from P1 and P2, including Spl, CRE, CPl, AP-2, NF-1, NF-kappa B, and C/EBP. Supershift assays using antibodies against C/EBP alpha and C/EBP beta and mutational analyses indicate that the protein binding to the C/EBP consensus recognition site at -925 to -933 is C/EBP alpha. The activity of promoter/CAT constructs containing the C/EBP recognition site is significantly decreased by cotransfection of C/EBP alpha but not C/EBP alpha but not C/EBP beta into either DDT1 MF-2 cells or primary rat hepatocytes. Partial hepatectomy causes a transient decrease in C/EBP alpha, as measured by DMSA, and an increase in beta 2 AR mRNA levels and rate of transcription in the remnant liver. Thus, derepression via C/EBP alpha is likely involved in the up-regulation of beta 2AR in the regenerating rat liver.

Authors+Show Affiliations

Department of Pharmacology & Toxicology, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8855951

Citation

Jiang, L, et al. "DNA Elements and Protein Factors Involved in the Transcription of the Beta 2-adrenergic Receptor Gene in Rat Liver. the Negative Regulatory Role of C/EBP Alpha." Biochemistry, vol. 35, no. 40, 1996, pp. 13136-46.
Jiang L, Gao B, Kunos G. DNA elements and protein factors involved in the transcription of the beta 2-adrenergic receptor gene in rat liver. The negative regulatory role of C/EBP alpha. Biochemistry. 1996;35(40):13136-46.
Jiang, L., Gao, B., & Kunos, G. (1996). DNA elements and protein factors involved in the transcription of the beta 2-adrenergic receptor gene in rat liver. The negative regulatory role of C/EBP alpha. Biochemistry, 35(40), 13136-46.
Jiang L, Gao B, Kunos G. DNA Elements and Protein Factors Involved in the Transcription of the Beta 2-adrenergic Receptor Gene in Rat Liver. the Negative Regulatory Role of C/EBP Alpha. Biochemistry. 1996 Oct 8;35(40):13136-46. PubMed PMID: 8855951.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - DNA elements and protein factors involved in the transcription of the beta 2-adrenergic receptor gene in rat liver. The negative regulatory role of C/EBP alpha. AU - Jiang,L, AU - Gao,B, AU - Kunos,G, PY - 1996/10/8/pubmed PY - 1996/10/8/medline PY - 1996/10/8/entrez SP - 13136 EP - 46 JF - Biochemistry JO - Biochemistry VL - 35 IS - 40 N2 - Primer extension and RNase protection analyses of the rat beta 2-adrenergic receptor (beta 2AR) gene identify two transcription start points at -64 and -220 nt, respectively. Transient transfections of putative promoter/pCAT constructs into DDT1 MF-2 cells indicate that fragments -36 to -100 (PI) and -186 to -312 (P2) are sufficient to promote transcription, whereas -911 to -1122 contains a negative regulatory element(s). RNase protection analysis of the 3' untranslated region (3'-UTR) indicates the presence of two transcripts with 3'-UTR of 111 and 604 nt exclusive of the poly(A+) tails. Northern blots of beta 2AR mRNA using full-length and partial cDNA probes indicate that a major 2.2 kb and a minor 1.6 kb species arise from the use of alternative promoters as well as different polyadenylation signals. DNase I footprinting and DNA mobility shift assays (DMSA) using rat liver nuclear extracts identify a number of transcription factors binding to sequence elements within or upstream from P1 and P2, including Spl, CRE, CPl, AP-2, NF-1, NF-kappa B, and C/EBP. Supershift assays using antibodies against C/EBP alpha and C/EBP beta and mutational analyses indicate that the protein binding to the C/EBP consensus recognition site at -925 to -933 is C/EBP alpha. The activity of promoter/CAT constructs containing the C/EBP recognition site is significantly decreased by cotransfection of C/EBP alpha but not C/EBP alpha but not C/EBP beta into either DDT1 MF-2 cells or primary rat hepatocytes. Partial hepatectomy causes a transient decrease in C/EBP alpha, as measured by DMSA, and an increase in beta 2 AR mRNA levels and rate of transcription in the remnant liver. Thus, derepression via C/EBP alpha is likely involved in the up-regulation of beta 2AR in the regenerating rat liver. SN - 0006-2960 UR - https://www.unboundmedicine.com/medline/citation/8855951/DNA_elements_and_protein_factors_involved_in_the_transcription_of_the_beta_2_adrenergic_receptor_gene_in_rat_liver__The_negative_regulatory_role_of_C/EBP_alpha_ L2 - https://doi.org/10.1021/bi960844o DB - PRIME DP - Unbound Medicine ER -